[1]巩天晓)),刘红涛),鲁照明),等.Notch1NICD胞内结构域真核表达载体的构建及其初步功能分析*[J].郑州大学学报(医学版),2009,(03):468-471.
 GONG Tianxiao)),LIU Hongtao),LU Zhaoming),et al.Construction of eukaryotic expression vectors of Notch1NICD and its preliminary functional study in esophageal squamous cell carcinoma cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(03):468-471.
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Notch1NICD胞内结构域真核表达载体的构建及其初步功能分析* ()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年03期
页码:
468-471
栏目:
食管癌研究
出版日期:
2009-05-01

文章信息/Info

Title:
Construction of eukaryotic expression vectors of Notch1NICD and its preliminary functional study in esophageal squamous cell carcinoma cells
作者:
巩天晓1)2)刘红涛3)鲁照明1)许培荣3)薛乐勋13)#
1)郑州大学第一附属医院细胞生物学研究室郑州4500522)郑州大学第二附属医院肿瘤科郑州4500143)郑州大学生物工程系细胞生物学研究室郑州450001
Author(s):
GONG Tianxiao1)2)LIU Hongtao3)LU Zhaoming1)XU Peirong3)XUE Lexun1)3)
关键词:
Notch1信号途径食管鳞状细胞癌细胞细胞增殖真核表达载体
Keywords:
Notch1 signaling pathway esophageal squamous cell carcinoma cell cell proliferation eukaryotic expression vector
分类号:
R735.1
摘要:
构建包含Notch1胞内结构域的Notch1NICD真核表达载体及绿色荧光蛋白载体并转染食管鳞状细胞癌细胞株EC9706及Eca109,并对Notch1NICD1在食管鳞状细胞癌中的功能进行初步探索。方法:根据GenBank上的Notch1NICD序列设计扩增引物,由RTPCR扩增NICD片段,测序鉴定,并通过Blast进行序列比对,然后回收目的片段并与真核表达载体pcDNA3.1(+)相连,构建Notch1NICD真核表达载体pNotch1NICD1,同时将扩增的NICD与pEGFPC1载体相连构建绿色荧光蛋白表达载体pENotch1NICD1,并转染EC9706及Eca109,观察Notch1在细胞中的定位及其途径的激活状态。同时将pNotch1NICD1真核表达载体转染EC9706及Eca109,用MTT法分别在24 h、48 h、72 h、96 h及120 h观察外源性Notch1片段的导入对EC9706及Eca109细胞增殖的影响。结果:通过RTPCR获得1个Notch1胞内区片段,并成功构建了包含Notch1胞内区结构域的绿色荧光蛋白载体及真核表达载体。转染pENotch1NICD1后,Notch1在胞质和胞核内均有表达,表明Notch1信号通路被激活。此外,MTT结果表明转染Notch1NICD1片段的食管鳞状细胞癌细胞的增殖受到明显抑制(P<0.05)。

参考文献/References:

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备注/Memo

备注/Memo:
*教育部“十五”“211工程”重点学科建设项目20022;河南省医学科技攻关计划项目20050009#通讯作者,男,1942年生,教授,研究方向:肿瘤分子生物学,Email:xuelx@371.net
更新日期/Last Update: 2010-05-28